Glutaryl-CoA Dehydrogenase Misfolding in Glutaric Acidemia Type 1

TY - JOUR

T1 - Glutaryl-CoA Dehydrogenase Misfolding in Glutaric Acidemia Type 1

AU - Barroso, Madalena

AU - Gertzen, Marcus

AU - Puchwein-Schwepcke, Alexandra F

AU - Preisler, Heike

AU - Sturm, Andreas

AU - Reiss, Dunja D

AU - Danecka, Marta K

AU - Muntau, Ania C

AU - Gersting, Søren W

PY - 2023/8/24

Y1 - 2023/8/24

N2 - Glutaric acidemia type 1 (GA1) is a neurotoxic metabolic disorder due to glutaryl-CoA dehydrogenase (GCDH) deficiency. The high number of missense variants associated with the disease and their impact on GCDH activity suggest that disturbed protein conformation can affect the biochemical phenotype. We aimed to elucidate the molecular basis of protein loss of function in GA1 by performing a parallel analysis in a large panel of GCDH missense variants using different biochemical and biophysical methodologies. Thirteen GCDH variants were investigated in regard to protein stability, hydrophobicity, oligomerization, aggregation, and activity. An altered oligomerization, loss of protein stability and solubility, as well as an augmented susceptibility to aggregation were observed. GA1 variants led to a loss of enzymatic activity, particularly when present at the N-terminal domain. The reduced cellular activity was associated with loss of tetramerization. Our results also suggest a correlation between variant sequence location and cellular protein stability (p < 0.05), with a more pronounced loss of protein observed with variant proximity to the N-terminus. The broad panel of variant-mediated conformational changes of the GCDH protein supports the classification of GA1 as a protein-misfolding disorder. This work supports research toward new therapeutic strategies that target this molecular disease phenotype.

AB - Glutaric acidemia type 1 (GA1) is a neurotoxic metabolic disorder due to glutaryl-CoA dehydrogenase (GCDH) deficiency. The high number of missense variants associated with the disease and their impact on GCDH activity suggest that disturbed protein conformation can affect the biochemical phenotype. We aimed to elucidate the molecular basis of protein loss of function in GA1 by performing a parallel analysis in a large panel of GCDH missense variants using different biochemical and biophysical methodologies. Thirteen GCDH variants were investigated in regard to protein stability, hydrophobicity, oligomerization, aggregation, and activity. An altered oligomerization, loss of protein stability and solubility, as well as an augmented susceptibility to aggregation were observed. GA1 variants led to a loss of enzymatic activity, particularly when present at the N-terminal domain. The reduced cellular activity was associated with loss of tetramerization. Our results also suggest a correlation between variant sequence location and cellular protein stability (p < 0.05), with a more pronounced loss of protein observed with variant proximity to the N-terminus. The broad panel of variant-mediated conformational changes of the GCDH protein supports the classification of GA1 as a protein-misfolding disorder. This work supports research toward new therapeutic strategies that target this molecular disease phenotype.

KW - Glutaryl-CoA Dehydrogenase/chemistry

KW - Amino Acid Metabolism, Inborn Errors/enzymology

KW - Brain Diseases, Metabolic/enzymology

KW - Protein Folding

KW - Mutation, Missense

KW - Protein Domains

KW - Humans

KW - Enzyme Stability

KW - Solubility

U2 - 10.3390/ijms241713158

DO - 10.3390/ijms241713158

M3 - SCORING: Journal article

C2 - 37685964

VL - 24

JO - INT J MOL SCI

JF - INT J MOL SCI

SN - 1661-6596

IS - 17

M1 - 13158

ER -